免疫磁球性能影响因素

Factors Effected on Performance of Immunomagnetic Beads for Bacteria Enrichment

DOI：10.3969/j.issn.1673-1689.2015.05.009

中文关键词: 免疫磁球 捕获率 磁分离时间 特异性 敏感性

英文关键词: immunomagnetic beads(IMB),capture rate,magnetic separation time,specificity,sensitivity

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中文摘要:

利用化学键法制备沙门氏菌捕获富集用免疫磁球,通过平板培养计数法确定其对目标菌的捕获率,对影响免疫磁球性能的磁分离时间、目标菌捕获率、特异性及敏感性等主要因素进行了研究。结果表明,免疫磁球在不同的磁分离时间内对目标菌的捕获率不同,在一定的的磁分离时间内捕获率达到最高；磁球与抗体的投料比影响免疫磁球的捕获性能,磁球200 μg、抗体25 μg的投料制备的免疫磁球能获得40%以上的捕获效率,具有一定的灵敏性(100 cfu/mL的目标菌捕获率为50%以上)；免疫磁球的粒径影响其捕获性能,大粒径(1 000 nm)免疫磁球目标菌捕获率小于小粒径(180 nm)免疫磁球目标菌捕获率；免疫磁球对单增李斯特菌、金黄色葡萄球菌、志贺氏菌非特异性在10%以下,在一定程度上影响免疫磁球的特异性捕获。

英文摘要:

In this study,we aims to study the main factors such as magnetic separation time,target capture rate,specificity,sensitivity that affect the performance of immunomagnetic beads (IMB). IMB were prepared chemically and the capture rate of target bacteria were determined by plate culture counting. The target bacteria capture rate varies in different magnetic separation time,which affected the capture rate,and the capture rate of IMB would reach the highest level. The amount of magnetic balls and antibody affect the capture performance of IMB. When the Feed ratio was 200 μg of magnetic ball and 25 μg of antibody,the capture rate would get more than 50%And IMB has a sensitivity (the capture rate is above 50% on target bacteria with 100 cfu /mL).The target capture rate of IMB with the large diameter(1 000 nm)is less than that of those with the small diameter (180 nm)at the same mass. The nonspecific capture rate for Escherichia coli,Staphylococcus aureusand Shigella was below 10%,which affected the immune specificity of IMB. This study provided a foundation and some data for application of IMB and detection of microorganisms.

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